Jiang Kun, Zhang Junhong, Ji Mingxia, Gai Pengzhou, Lv Qiaoying
Department of Orthopedics, The Affiliated Yantai Yuhuangding Hospital of Qingdao University, Yantai 264000, P.R. China.
J BUON. 2019 Jul-Aug;24(4):1706-1711.
To evaluate the inhibitory effect of 5-fluorouracil (5-FU) in combination with cisplatin on the proliferation of human osteosarcoma cells in vitro.
Three groups of human osteosarcoma U2OS cells were cultured in DMEM medium supplemented with the following drugs: 20 µg/mL 5-FU, 50 µg/mL 5-FU and 50 µg/mL 5-FU in combination with 0.5 mg/L cisplatin. After culture at 24, 48 and 72 h, the inhibition of proliferation rate of U2OS cells was calculated by CCK-8 cell kit. Cell invasiveness was assessed by Transwell assay. Flow cytometry was used for monitoring the cell apoptosis.
5-FU inhibited the growth of osteosarcoma cells, and the results of different concentrations of 5-FU were significantly different. The growth of U2OS cells decreased significantly within 24-72 h, and the concentration of 5-FU increased with time. The inhibition of the shift was more obvious, and the combined drug inhibition was significantly higher than the 20μg/ml 5-FU Group, 0.5mg/L Cisplatin Group and 50μg/ml 5-FU Group. After 72 h, the mean inhibitory rates of 20 μg/mL 5-FU, 50 μg/mL 5-FU, 50 μg/ml 5-FU in combination with 0.5 mg/L cisplatin, and 0.5 mg/L cisplatin were 12.54±1.26%, 22.17±0.59%, 32.54±1.25%, 20.84±0.83% respectively, and the difference was significant (p<0.05). Results of cell invasion assay showed that after culturing for 48 h, the mean number of cells penetrating the membrane was 22.84±5.27 in the culture group of 0.5 mg/L cisplatin, 30.57±5.68 in the culture group of 20 µg/mL 5-FU, 18.68±4.88 in the culture group of 50 µg/mL 5-FU, and 9.84±3.64 in the culture group of 50 µg/mL 5-FU in combination with 0.5 mg/L cisplatin, respectively, and 72.00±7.52 in the control group, showing statistical differences in each group (p<0.05). The apoptosis of the control group was significantly lower than that of the other groups (p<0.05). Apoptosis rate of the 20μg/mL 5-FU group was significantly lower than that of the 0.5mg/L cisplatin group, the 50μg/mL 5-FU group and the 50 μg/ml 5-FU group in combination with 0.5mg/L cisplatin (p<0.05). There was no difference in apoptosis between 0.5mg/L cisplatin and 50μg/mL 5-FU group (p>0.05).
5-FU in combination with cisplatin exerts an inhibitory effect on the proliferation and invasion of human osteosarcoma cells in vitro, and can promote cell apoptosis.
评估5-氟尿嘧啶(5-FU)联合顺铂对人骨肉瘤细胞体外增殖的抑制作用。
将三组人骨肉瘤U2OS细胞培养于添加以下药物的DMEM培养基中:20μg/mL 5-FU、50μg/mL 5-FU以及50μg/mL 5-FU与0.5mg/L顺铂联合使用。在24、48和72小时培养后,通过CCK-8细胞试剂盒计算U2OS细胞增殖率的抑制情况。通过Transwell实验评估细胞侵袭能力。采用流式细胞术监测细胞凋亡情况。
5-FU抑制骨肉瘤细胞生长,不同浓度的5-FU结果差异显著。U2OS细胞在24 - 72小时内生长显著下降,且5-FU浓度随时间增加。联合用药组的抑制作用更明显,并显著高于20μg/ml 5-FU组、0.5mg/L顺铂组和50μg/ml 5-FU组。72小时后,20μg/mL 5-FU、50μg/mL 5-FU、50μg/ml 5-FU与0.5mg/L顺铂联合使用组以及0.5mg/L顺铂组的平均抑制率分别为12.54±1.26%、22.17±0.59%、32.54±1.25%、20.84±0.83%,差异有统计学意义(p<0.05)。细胞侵袭实验结果显示,培养48小时后,0.5mg/L顺铂培养组穿透膜的细胞平均数量为22.84±5.27,20μg/mL 5-FU培养组为30.57±5.68,50μg/mL 5-FU培养组为18.68±4.88,50μg/ml 5-FU与0.5mg/L顺铂联合使用培养组为9.84±3.64,对照组为72.00±7.52,各组间差异有统计学意义(p<0.05)。对照组的凋亡率显著低于其他组(p<0.05)。20μg/mL 5-FU组的凋亡率显著低于0.5mg/L顺铂组、50μg/mL 5-FU组以及50μg/ml 5-FU与0.5mg/L顺铂联合使用组(p<0.05)。0.5mg/L顺铂组与50μg/mL 5-FU组之间的凋亡情况无差异(p>0.05)。
5-FU联合顺铂对人骨肉瘤细胞的体外增殖和侵袭具有抑制作用,并可促进细胞凋亡。